Repetitive Synthesis of High‐Molecular‐Weight Hyaluronic Acid with Immobilized Enzyme Cascades

Abstract

Industrial hyaluronic acid (HA) production comprises either fermentation with Streptococcus strains or extraction from rooster combs. The hard‐to‐control product quality is an obstacle to these processes. Enzymatic syntheses of HA were developed to produce high‐molecular‐weight HA with low dispersity. To facilitate enzyme recovery and biocatalyst re‐use, here the immobilization of cascade enzymes onto magnetic beads was used for the synthesis of uridine‐5’‐diphosphate‐α‐d‐N‐acetyl‐glucosamine (UDP‐GlcNAc), UDP‐glucuronic acid (UDP‐GlcA), and HA. The combination of six enzymes in the UDP‐sugar cascades with integrated adenosine‐5'‐triphosphate‐regeneration reached yields between 60 and 100 % for 5 repetitive batches, proving the productivity. Immobilized HA synthase from Pasteurella multocida produced HA in repetitive batches for three days. Combining all seven immobilized enzymes in a one‐pot synthesis, HA production was demonstrated for three days with a HA concentration of up to 0.37 g L−1, an average MW of 2.7–3.6 MDa, and a dispersity of 1.02–1.03.

Polysaccharide-Based Transdermal Drug Delivery

“…HA is a linear glycosaminoglycan (GAG) consisting of N -acetyl- d -glucosamine and d -glucuronic acid. HA derived from rooster comb [ 41 ], microbial source [ 5 ]. HA was first isolated from the vitreous humor of bovine eyes.…”

Prospective bacterial and fungal sources of hyaluronic acid: A review

“…This approach is especially useful for obtaining non-natural chemicals [106] , [107] . Recent studies have shown a high efficiency of using enzyme cascades immobilized by magnetic beads for obtaining HW HA [108] . Several reviews on strategies for chemoenzymatic synthesis with an emphasis on the use of enzyme cascades have been published recently [106] , [109] .…”

One‐Step Preparation of Cell‐Free ATP Regeneration Module Based on Non‐Oxidative Glycolysis Using Thermophilic Enzymes

“…[10] Moreover, the accumulation of low length polyP and orthophosphate in the reaction mixture is not favorable, especially for repetitive batch reactions, because of their tendency to form insoluble salts with divalent metal ions. [13,14] One promising approach is the in vitro reconstitution and implementation of non-oxidative glycolysis (NOG). The NOG pathway can fully produce stoichiometric metabolites from dextrose sources to C2 metabolites, without any carbon loss.…”

“…Despite its simple approach, PPK is typically expressed as an inclusion body [12] and its activity depends on the length of the polyP chain, which can differ greatly depending on the suppliers and their respective production methods [10] . Moreover, the accumulation of low length polyP and orthophosphate in the reaction mixture is not favorable, especially for repetitive batch reactions, because of their tendency to form insoluble salts with divalent metal ions [13,14] …”